Rapid detection of functional expression of C-5-DNA methyltransferases in yeast.
We have previously employed the cytosine-5-DNA methyltransferase (MTase), M. Sss I, as a probe for chromatin architecture in intact cells. Although M. Sss I offers the highest resolution of any currently available MTase, the difficulty in establishing stable, methylation-positive strains poses a barrier to its general utility as a chromatin probe. We describe a simple screen for M. Sss I-expressing strains that eliminates the purification of PCR products amplified from bisulfite-treated DNA, use of radioisotopes, polyacrylamide sequencing gel electrophoresis, and autoradiography. The high throughput of the method now makes it feasible to introduce M. Sss I into a variety of wild-type and mutant genetic backgrounds.
Kladde
M P
MP
Department of Biochemistry and Molecular Biology and Center for Gene Regulation, The Pennsylvania State University, 308 Althouse Laboratory, University Park, PA 16802-4500, USA. mpk6@psu.edu
Simpson
R T
RT
D001483:Base Sequence; D002843:Chromatin / Q000235:genetics; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics*; D019175:DNA Methylation; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D009154:Mutation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*
D002843:Chromatin; D017931:DNA Primers; D004271:DNA, Fungal; D004248:DNA (Cytosine-5-)-Methyltransferases